Antibiotic-resistant bacteria pose a substantial health risk. The available methods to detect resistant strains are expensive, time-consuming, and complicated. Nomadics will functionalize a highly sensitive amplifying fluorescent polymer (AFP) to detect extremely low concentrations of bacterial antibiotic-resistance markers. The proposed system will provide comparable or improved sensitivity, with real-time detection in an easy-to-use, low-cost device that will support point-of-care identification of resistant pathogens. This novel detection system uses the intrinsic amplification of fluorescence transduction events afforded by AFP and will detect the mecA gene and PBP2a protein, indicators of resistance in methicillin-resistant Staphylococcus aureus. Specific probe sequences and monoclonal antibodies will provide the target recognition event, which will result in signal amplification by AFP and detection by the sensor. The sensitivity, specificity, and rapidity of the system will be tested in vitro and in vivo using methicillin-resistant, borderline resistant, and methicillin-sensitive S. aureus strains as well as other staphylococcal and bacterial species and will be compared to other resistance detection technologies. Ultimately, the proposed research will lead to commercial production of a sensitive, rapid, affordable, point-of-care detector capable of identifying a variety of resistant bacteria in clinical samples, thus permitting immediate selection of effective antibiotic therapy. PROPOSED COMMERCIAL APPLICATION: The proposed technology will be used to determine whether a Staphylococus aureus infection is resistant to oxacillin. However, the technology will be applicable to detection of other antimicrobial resistance genes and proteins, as well as other microorganisms. This capability will allow the immediate use of appropriate antibiotics to fight infection without increasing resistance to other antibiotics. Additionally, the method proposed can be used to identify other chemical properties that are useful in identifying organisms. The technology is simple, which will allow it to be used outside the laboratory by minimally skilled individuals. As a low-cost technology, the detection system will find widespread use in point-of-care facilities, in field applications, and in under-developed regions.